Molecular and General Genetics MGG

, Volume 242, Issue 2, pp 121–129 | Cite as

Cloning of the blasticidin S deaminase gene (BSD) from Aspergillus terreus and its use as a selectable marker for Schizosaccharomyces pombe and Pyricularia oryzae

  • Makoto Kimura
  • Takashi Kamakura
  • Quan Zhou Tao
  • Isao Kaneko
  • Isamu Yamaguchi
Original Articles


Aspergillus terreus produces a unique enzyme, blasticidin S deaminase, which catalyzes the deamination of blasticidin S (BS), and in consequence confers high resistance to the antibiotic. A cDNA clone derived from the structural gene for BS deaminase (BSD) was isolated by transforming Escherichia coli with an Aspergillus cDNA expression library and directly selecting for the ability to grow in the presence of the antibiotic. The complete nucleotide sequene of BSD was determined and proved to contain an open reading frame of 393 bp, encoding a polypeptide of 130 amino acids. Comparison of its nulceotide sequence with that of bsr, the BS deaminase gene isolated from Bacillus cereus, indicated no homology and a large difference in codon usage. The activity of BSD expressed in E. coli was easily quantified by an assay based on spectrophotometric recording. The BSD gene was placed in a shuttle vector for Schizosaccharomyces pombe, downstream of the SV40 early region promoter, and this allowed direct selection with BS at high frequency, following transformation into the yeast. The BSD gene was also employed as a selectable marker for Pyricularia oryzae, which could not be transformed to BS resistance by bsr. These results promise that the BSD gene will be useful as a new dominant selectable marker for eukaryotes.

Key words

Aspergillus terreus Blasticidin S deaminase gene Selectable marker Schizosaccharomyces pombe Pyricularia oryzae 


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Copyright information

© Springer-Verlag 1994

Authors and Affiliations

  • Makoto Kimura
    • 1
  • Takashi Kamakura
    • 2
  • Quan Zhou Tao
    • 1
  • Isao Kaneko
    • 1
  • Isamu Yamaguchi
    • 1
  1. 1.Microbial Toxicology LaboratoryInstitute of Physcial and Chemical Research (RIKEN)Wako-shi, SaitamaJapan
  2. 2.Biodesign Research GroupInstitute of Physical and Chemical Research (RIKEN)Wako-shi, SaitamaJapan

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